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Detection of marker-free precision genome editing and genetic variation through the capture of genomic signatures

Genome editing technologies have transformed our ability to engineer desired genomic changes within living systems. However, detecting precise genomic modifications often requires sophisticated, expensive, and time-consuming experimental approaches. Here, we describe DTECT (Dinucleotide signaTurE Ca...

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Main Authors: Billon, Pierre (Author) , Nambiar, Tarun S. (Author) , Hayward, Samuel B. (Author) , Zafra, Maria P. (Author) , Schatoff, Emma M. (Author) , Oshima, Koichi (Author) , Dunbar, Andrew (Author) , Breinig, Marco (Author) , Park, Young C. (Author) , Ryu, Han S. (Author) , Tschaharganeh, Darjus-Felix (Author) , Levine, Ross L. (Author) , Baer, Richard (Author) , Ferrando, Adolfo (Author) , Dow, Lukas E. (Author) , Ciccia, Alberto (Author)
Format: Article (Journal)
Language:English
Published: March 10, 2020
In: Cell reports
Year: 2020, Volume: 30, Issue: 10, Pages: 3280-3295,e1-e6
ISSN:2211-1247
DOI:10.1016/j.celrep.2020.02.068
Online Access:lizenzpflichtig
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Author Notes:Pierre Billon, Tarun S. Nambiar, Samuel B. Hayward, Maria P. Zafra, Emma M. Schatoff, Koichi Oshima, Andrew Dunbar, Marco Breinig, Young C. Park, Han S. Ryu, Darjus F. Tschaharganeh, Ross L. Levine, Richard Baer, Adolfo Ferrando, Lukas E. Dow, and Alberto Ciccia
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Summary:Genome editing technologies have transformed our ability to engineer desired genomic changes within living systems. However, detecting precise genomic modifications often requires sophisticated, expensive, and time-consuming experimental approaches. Here, we describe DTECT (Dinucleotide signaTurE CapTure), a rapid and versatile detection method that relies on the capture of targeted dinucleotide signatures resulting from the digestion of genomic DNA amplicons by the type IIS restriction enzyme AcuI. DTECT enables the accurate quantification of marker-free precision genome editing events introduced by CRISPR-dependent homology-directed repair, base editing, or prime editing in various biological systems, such as mammalian cell lines, organoids, and tissues. Furthermore, DTECT allows the identification of oncogenic mutations in cancer mouse models, patient-derived xenografts, and human cancer patient samples. The ease, speed, and cost efficiency by which DTECT identifies genomic signatures should facilitate the generation of marker-free cellular and animal models of human disease and expedite the detection of human pathogenic variants.
Item Description:Gesehen am 30.04.2020
Physical Description:Online Resource
ISSN:2211-1247
DOI:10.1016/j.celrep.2020.02.068

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